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UTERUSUTERUSEXPRESSION OF INTERLEULIN 11 AND ITS RECEPTOR IN THE HUMANENDOMETRIUM THROUGHOUT THE MENSTRUAL CYCLE.Evdokia Dimitriadis, Lorraine Robb l , Lois A SalamonsenPrince Henry's Institute ofMedical Research, P.O. Box 5152, Clayton, 3168,Victoria, Australia'Walter and Eliza Hall Institute ofMedical Research, Parkville, 3050, Victoria, AustraliaIntroductionInterleukin 11 (lL-II) is a cytokine with pleitropicactivities on a range of cell types (1). IL-IIsignals via a homodimeric receptor complexcomposed of a specific binding chain, IL-llreceptor alpha (lL-IIRD) and the signallingcomponent gp 130. IL-ll plays a crucial role infemale fertility in the mouse (2). Female micewith a null mutation of the IL-l1RD chain wereinfertile due to a defective decidualizationresponse of the uterine stroma to the implantingblastocyst. In the human female, decidualizationof the stromal cells occurs in the secretory phaseof the menstrual cycle and involves biochemicaland morphological changes. If implantationoccurs, stromal cell decidualization continuesthroughout pregnancy. Therefore factors involvedin· decidualization are important for implantationand subsequent placentation. As a first step inelucidating the role of IL-llin human fertility theaim of this study was to examine the localizationand expression pattern of IL-l1 in the humanendometrium throughout the menstrual cycle.MethodsEndometrial biopsies were obtained from 30women who showed no apparent endometrialdysfunction. Biopsy sections were stained byimmunohistochemistry to immunolocalise IL-ll.Menstrual cycle sections were divided into 6phases with the following day ranges:Menstrual (ME) d1-5, early-proliferative (EP) d5­8, late-proliferative (LP) d9-13, early-(ES) d 14-18mid-eMS) d19-23 and late-(LS) d24-28 secretory,consisting of4 samples per phase.Antisera specific for human recombinant IL-l 1(R&D Systems) was used forimmunohistochemistry. Semi-quantitative scoringof staining intensity and distribution wasperformed. Results are expressed as means. Theexpression of IL-l1 transcripts in late secretoryphase endometrium was determined by in situhybridisation and IL-llR[ : mRNA expressionacross the menstrual cycle by reversetranscriptase-polymerase chain reaction (RT­PCR).ResultsImmunoreactive IL-ll was found associated withall cellular compartments. In the ME, and LPphases glandular epithelial and stromal cellsstained positively (Figure 1). In contrast, insecretory phase tissue, staining was seen in allcellular compartments (Figure 1). In the ES phasemoderate immunoreactivity was found in theluminal (LE) and glandular epithelial (GE) cells.More intense staining was seen in the endothelial(endo) and vascular smooth muscle (VSM) cellsin .the MS phase. However the most intensestaining was seen in the decidual cells of the LSphase, when they were present, with the nondecidualizedstromal cell staining intensityvarying little in the MS and LS phases. Theseresults were verified by in situ hybridisation. TheIL-II Ra was also found to be present throughoutthe cycle.Figure 1LEGEslromaendaVSM[REP-I"ICLPI DEs!111.151j ll:lLS I~decidualConclusionsImmunoreactive IL-ll is found in the humanendometrium in all the major cell types.IL-ll Ra mRNA was present throughout the cycle.In the proliferative phase moderate IL-Il stainingoccured in the glandular epithelial and stromalcells.Late secretory phase endometrial tissue showedthe most intense staining for IL-II which wasverified by in situ hybridisation.The most intense staining observed occurred inthe decidual cells.The presence of in vivo IL-II in a normalphysiological situation in secretory phaseendometrium implies that IL-II may be involvedin decidualization and the preparation of areceptive endometrium for implantation.ReferencesDu et al (1997) Blood 89: 3897-3908Robb et al (1998) Nature Med 4: 303-308INTRAUTERINE SEMINAL PLASMA INDUCES ENDOMETRIALINFLAMMATORY RESPONSE IN GILTSS. O'Leary, D.T. Annstrong, R Kamai and S.A. RobertsonDepartment ofObstetrics and Gynaecology, The University ofAdelaide, Adelaide, SA 5005IntroductionSeminal plasma (SP) has previously been thought tofunction simply as a transport and swvival mediumfor spermatozoa traversing the female reproductivetract However, emerging data in rodents nowindicates that factors in SP, most notablytransfonning growth factor beta (TGF~), act toevoke an inflammatory response in the uterineendometrium after insemination (1). This responseapPearS to improve implantation through inducingembryotrophic cytokine. expression and promotingimmunological tolerance to paternal transplantationantigens expressed on the semi-allogeneic conceptus.In pigs, exposure to seminal constituents has beenshown to improve litter size (2) and to influenceovarian function (3,4), but the physiologicalmechanisms mediating this effect are unknown. Theaim of this study was to begin to investigate themolecular basis of the interaction between SP andthe female reproductive tract in pigs. Specifically,we have measured the acute effect of SP onleukocyte recruitment and other inflammatoryparameters in the uteri ofgilts.Materials and MethodsAt the onset ofgonadotrophin-induced oestrus, eightLarge White X Landrace gilts were administeredeither seminal plasma (SP; n=4) or saline (pBS~ n=4)by transcervical infusion~ Reproductive tracts wereretrieved at autopsy 36 hours after treatment whenthe weight, luminal fluid content and vascular indexof uteri were scored. Endometrial tissue biopsieswere frozen in ocr and the abundance and locationof resident leukocytes were assessed byimmunohistochemical analysis of ethanol-fixedfrozen sections using monoclvnal antibodies specificfor CD45 (MCAI222), :MIlC class II (MSA3), andmacrophages (HB142). Antibody reactivity (%positivity) was quantified by video image analysis.Fig. 1. The. effect of intrauterine SP infusion onleukocyte populations in the endometrium of gilts-'-- .80 -A~70f 60t1 50l:lot'$. 4090..,--------------A'1-. .-1- At II30 T • • PBS20 •A SP10it = P